Isolation and structure elucidation of highly antioxidative 3,8″-linked biflavanones and flavanone-C-glycosides from Garcinia buchananii bark

The aim of this study was to identify antioxidants from Garcinia buchananii bark extract using hydrogen peroxide scavenging and oxygen radical absorbance capacity (ORAC) assays. LC-MS/MS analysis, 1D- and 2D-NMR, and circular dichroism (CD) spectroscopy led to the unequivocal identification of the major antioxidative molecules as a series of three 3,8″-linked biflavanones and two flavanone-C-glycosides. Besides the previously reported (2R,3R,2″R,3″R)-naringenin-C-3/C-8″ dihydroquercetin linked biflavanone (GB-2; 4) and (2R,3S,2″R,3″R)-manniflavanone (3), whose stereochemistry has been revised, the antioxidants identified for the first time in Garcinia buchananii were (2R,3R)-taxifolin-6-C-β-D-glucopyranoside (1), (2R,3R)-aromadendrin-6-C-β-D-glucopyranoside (2), and the new compound (2R,3S,2″S)-buchananiflavanone (5). The H?O? scavenging and the ORAC assays demonstrated that these natural products have an extraordinarily high antioxidative power, especially (2R,3S,2″R,3″R)-manniflavanone (3) and GB-2 (4), with EC?? values of 2.8 and 2.2 μM, respectively, and 13.73 and 12.10 μmol TE/ μmol. These findings demonstrate that G. buchananii bark extract is a rich natural source of antioxidants.     

Hydrolysis of Chicoric and Caftaric Acids with Esterases and Lactobacillus johnsonii in Vitro and in a Gastrointestinal Model

Chicoric acid (ChA) and caftaric acid (CafA) were identified as bioactive components of chicory and have been ascribed a number of health benefits. This study investigated the hydrolysis of ChA and CafA with enzymes and a probiotic bacterium Lactobacillus johnsonii (La1). Esterase from Aspergillus japonicus (24 U/mg) hydrolyzed 100% of ChA (5 mM) and CafA (5 mM) after 3 h, at pH 7.0 and 37 °C. Under the same reaction conditions, 100% hydrolysis of ChA and CafA was achieved with a spray-dried preparation of La1. The addition of La1 (100 mg/mL, 3.3 E9 cfu/g) to CafA solution in a gastrointestinal model (GI model) resulted in 65% hydrolysis of CafA. This model simulates the physicochemical conditions of the human gastrointestinal tract. No hydrolysis of CafA was observed after passage through the GI model in the absence of La1. The results of this study support the hypothesis that ChA and CafA are degraded by gut microflora before absorption and metabolization.     

Soil microbial biomass and its activity estimated by kinetic respiration analysis – Statistical guidelines

The use of kinetic respiration analysis to determine soil microbial biomass its active part and maximum specific growth rate has recently increased. With this method, the increase in soil respiration rate initiated by application of carbon growth substrate, e.g. glucose, and mineral nutrients is used to estimate parameters describing microbial growth in soil. This study refines the method by developing statistical guidelines for the data analysis and processing. The kinetic respiration analysis assumes that microbial growth is not limited by substrate and energy. That is why it is critically important to identify the time period corresponding to the unlimited growth. In this work, we studied how the unlimited growth phase can be identified in less subjective ways by examining 121 datasets of respiration time series of 44 different soil samples taken from field plots. Deflection of the respiratory curve from the exponential pattern indicates growth limitation. Subjective selection of the part of respiratory curve which fits to the exponential pattern resulted in a 30% bias in specific microbial growth rates. We propose rules that are based on inspecting the patterns in a series of plots of residuals of fitted respiration rate. By comparing those rules with a set of statistical criteria we find that the weighted-coefficient of determination (r²) can be used to objectively constrain the unlimited growth phase in those cases where double-limitation does not occur. Furthermore, we discuss how the uncertainty of estimated microbial parameters is influenced by a) measurement uncertainty, b) biased measurement at the beginning of the experiment, and c) the number and timing of respiration measurements. We recommend checking plots of fits and residuals as well as reporting uncertainty bounds together with the estimated microbial parameters. A free statistical package is provided to easily deal with those aspects.     

Plant genetic resources of Lemnos (Greece), an isolated island in the Northern Aegean Sea, with emphasis on landraces

Lemnos Island is located in the Northern Aegean Sea and presents a rich biodiversity in plant genetic resources including wild species and crop landraces. Landraces have been cultivated under traditional agricultural systems and many have survived genetic erosion. They are mainly conserved in home gardens or in small fields, usually by elderly people, and are limited to local consumption. Two collecting expeditions were organized in Lemnos, a minor one in 2009 and a larger one in 2010. The results of the two current expeditions are discussed and compared with previous expeditions in the island. An overview is given on the crops cultivated, the landraces collected and those considered as lost, alongside with some information about their traditional uses. Finally, the importance of landraces in modern agriculture and preservation policies in Lemnos are discussed.     

Eradication of PVS from potato clones through excision of meristems from in vitro,heat-treated shoot tips

In vitro shoot tips from PVS-infected potato breeding clones and cultivars were exposed to a constant temperature of 34 C and 8,100 lux light. Meristems were excised from heat-treated shoot tips after 3 to 7 weeks of heat therapy. Serological tests were performed on shoot tips derived from meristems excised from heat-treated shoot tips. Additional serological tests, combined with bioassays, applied to six-week-old pot-grown cuttings derived from these shoot tips, did not detect PVS in 19 out of 21 clones given heat therapy. A 90% rate of eradication was achieved.     

THE VEGETATION OF THE PEWSEY VALE ESCARPMENT, WILTSHIRE

Records were made on a standard point-quadrat frame at intervals of 20 feet along fifteen transects down the chalk escarpment on the north side of Pewsey Vale in Wiltshire. These records demonstrate how great is the variability in chalk grassland, both in different parts of each transect and in corresponding parts of adjacent transects. Chalk-down vegetation must be regarded as unstable and greatly altered by past and present use of the land for cultivation and grazing.     

Economic injury levels for Bemisia tabaci (Homoptera: Aleyrodidae) in cotton: impact of crop price, control costs, and efficacy of control

Field studies were conducted during 1993 and 1994 in the Imperial Valley, California and Maricopa, Arizona to examine relationships between densities of Bemisia tabaci (Gennadius) populations and cotton (Gossypium hirsutum L.) yields, and to estimate economic injury levels (EILs) for pest management application. Populations of B. tabaci were manipulated by applying different numbers of insecticide applications in replicated plots. Resulting insect populations and cotton lint yields were used to develop damage functions and to estimate EILs for all life stages in relation to variable cotton prices, insecticide costs, and pest control efficacy. Economic injury levels declined with increasing cotton prices and increased as the cost of control increased, however, these changes were relatively small, based on typical ranges in price and control costs. In contrast, the efficacy of control provided by insecticide applications had a large influence on EILs, with lower efficacies being associated with higher injury levels. We developed a multiple regression model that accounted for the dynamic changes in the EIL in relation to crop price, control costs, control efficacy, and potential yield. Based on average prices and reasonable control costs and efficacy, EILs ranged from 5.9–15.2 adults/leaf, 6.1–19.8 eggs cm⁻², and 1.7–4.7 nymphs cm⁻² of leaf area. Additional research is needed to more closely relate the costs of control to the suppression of insect populations, and to define economic thresholds that will enable pest managers to maintain pest populations below EILs.     

Wound-induced suberization and periderm development in potato tubers as affected by temperature and gamma irradiation

Summary Histochemical evaluation of the wound-induced suberization and periderm formation, the processes of wound healing, in potato tubers (Solanum tuberosum L.) showed that both the processes occur most rapidly at 25°C. Wound-healing is delayed at 10 or 15°C while a temperature of 35°C prevented periderm formation and retarded suberization. Gamma irradiation up to 100 Gy, the optimal dose for sprout inhibition, did not affect suberization, which suggests that the DNA-replicating mechanism is more radiation-sensitive than suberin biosynthesis. A dose of 20 to 30 Gy, which had no effect on sprouting inhibited wound periderm formation indicating that meristems in resting buds are apparently less sensitive to irradiation than nuclei of the potential periderm cells. It seems probable that a major cause for the bacterial soft rot occurring in tubers when stored under high tropical ambient temperatures or when irradiated for sprout inhibition is due to an impairment of the wound periderm formation.

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Zusammenfassung Um die m?glichen Gründe für den gesteigerten mikrobiellen Abbau von Kartoffeln w?hrend der Lagerung unter tropischen Umweltbedingungen oder nach der Bestrahlung mit R?ntgenstrahlen zur Keimhemmung aufzukl?ren, wurde der Einfluss von Temperatur und Bestrahlung auf die durch Verletzung induzierte Suberin-und Peridermentwicklung untersucht. Es zeigte sich, dass sowohl die Suberinisierung als auch die Peridermentwicklung bei 25°C schneller verlaufen, w?hrend sie bei 10–15°C verlangsamt sind. Temperaturen über 35°C verhindern die Peridermbildung und verz?gern die Suberinisierung (Tabelle 1). Das Ausmass der Polyphenolbildung war im 1 mm unter der Verletzung bei 20, 25 und 28–30°C ?hnlich der bei 10°C und 15°C w?hrend es bei 35°C verringert war (Abb. 2). Histochemische Untersuchungen zeigten, dass die Suberinisierung in unbestrahlten Knollen und in Knollen, die bis zu 100 Gy erhalten hatten, der optimalen Dosis für die Keimhemmung, im gleichen Ausmass erfolgte (Tabelle 2). Aus diesen Ergebnissen wird geschlossen, dass der Verdopplungsmechanismus der DNS auf Bestrahlung empfindlicher reagiert als die Suberinsynthese. Der Gehalt an Polyphenolen in den suberinisierten Zellschichten war in bestrahlten Knollen vergleichsweise h?her (Tabelle 3). 20–30 Gy an R?ntgenstrahlen reichten aus, um die Wundperidermbildung zu unterbinden (Tabelle 2 und Abb. 1), w?hrend für die Keimhemmung eine Minimaldosis von 50 Gy ben?tigt wurde. Das bedeutet, dass die Meristeme in den ruhenden Knospen auf die Bestrahlung weniger empfindlich reagieren als die Kerne in den potentiellen Peridermzellen. Die suberinisierten Zellschichten k?nnen von der Wundoberfl?che unbestrahlter Knollenh?lften leicht abgeschoben werden, w?hrend das bei Knollen, die einer Bestrahlung von 20 Gy und mehr ausgesetzt waren, nicht m?glich ist. Dieser Unterschied k?nnte als Methode zur Identifizierung bestrahlter Knollen verwendet werden. Die Ergebnisse zeigen, dass die Sch?digung der Wundperidermentwicklung ein Hauptgrund für die gesteigerte mikrobielle F?ule von Kartoffelknollen w?hrend der Lagerung bei tropischen Temperaturen oder nach Bestrahlung zur Keimhemmung ist.

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Résumé L'influence de la température et de l'irradiation sur la subérisation d'une blessure occasionnée et sur le développement du périderme a été étudiée, dans le but d'expliquer les déchets de pourritures microbiennes sur des tubercules mis en conservation sous des conditions de climat tropical ou à la suite d'une exposition aux rayons gamma en vue de l'inhibition de la germination. Le développement du périderme et la subérisation se sont effectués rapidement à 25°C mais les réactions ont été retardées à 10 ou 15°C. Une température de 35°C a empêché la formation de périderme et retardé la subérisation (tableau 1). Le taux de polyphénol dans la 1ère couche d'1 mm sous la blessure était sensiblement le même à 10, 15, 20, 25 et 28–30°C mais il avait diminué à 35°C (figure 2). Les résultats de l'analyse hystochimique ont montré que la subérisation avait progressé à vitesse égale pour des tubercules non-irradiés et des tubercules irradiés à plus de 100 Gy, dose optimale correspondant à l'inhibition de la germination. Ces résultats suggèrent que le mécanisme de réplication de l'ADN est plus sensible à la radiation que la biosynthèse de la subérine. Le niveau de polyphénols dans les couches cellulaires en voie de subérisation était plus élevé dans les tubercules irradiés que dans les tubercules non irradiés (tableau 3). 20 à 30 Gy de rayons gamma ont suffi pour empêcher la formation du périderme de cicatrisation (tableau 2 et figure 1) tandis que la dose minimale nécessaire à l'inhibition de la germination était de 50 Gy. Les méristèmes de germes en dormance seraient donc moins sensibles à l'irradiation que les noyaux des cellules du périderme potentiel. Les couches cellulaires subérisées peuvent facilement se détacher de la surface endommagée lorsqu'il s'agit de tubercules non irradiés, tandis qu'elles sont difficilement séparables dans le cas de tubercules exposés à 20 Gy ou plus. Cette différence peut conduire à une méthode d'identification des tubercules irradiés. La recrudescence des pourritures microbiennes sur tubercules mis en conservation à des températures tropicales ou exposés aux rayons gamma en vue d'inhibition de la germination, semble liée principalement au ralentisement du développement d'un périderme de cicatrisation.

     

SHEEP PATHSO

The variability of some chalk pastures above the Pewsey Vale in Wiltshire was studied by recording point-quadrats two inches apart on strips across sheep paths; the results are shown diagrammatically. There were usually lines of tall grasses at the edges of the paths, with belts of shorter grass, richer in forbs, between the paths. Some species of grasses and forbs were most common on or near the paths; others were more common in the short turf. The theoretical and practical implications of this variability are briefly discussed.     

Vibrational circular-dichroism spectroscopy of homologous cyclic peptides designed to fold into β helices of opposite chirality

Cyclic β-helical peptides have been developed as model structured biomolecules for examining peptide adsorption and conformation on surfaces. As a key prerequisite to circular-dichroism (CD) analysis of these model peptides on surfaces, their conformations and the corresponding vibrational spectra in the 1400-1800 cm?1 range were analyzed by vibrational circular-dichroism (VCD) spectroscopy in solution. The two model peptides ("β Leu and β Val") were examined in chloroform, where they each fold into a homogeneous well-defined antiparallel double-stranded β-helical species, as determined previously by NMR and electronic CD spectroscopy. Because the β-helical conformations of β Leu and β Val are well characterized, the VCD spectra of these peptides can be unambiguously correlated with their structures. In addition, these two β-helical peptides differ from one another in two key respects that make them uniquely advantageous for CD analysis--first, while their backbone conformations are topologically similar, β Leu and β Val form helices of opposite chiralities; second, the two peptides differ in their sequences, i.e., composition of the side chains attached to the backbone. The observed VCD spectra for β Leu and β Val are roughly mirror images of each other, indicating that the VCD features are dominated by the chirality and conformation of the peptide backbone rather than by the peptide sequence. Accordingly, spectra similarly characteristic of peptide secondary structure can be expected for peptides designed to be structural analogs of β Leu and β Val while incorporating a variety of side chains for studies of surface adsorption from organic and aqueous solvents.